There are known many methods and devices for detecting and measuring of the biomedical agents (hereinbelow, the terms “biomedical agent” and/or “agent” and/or “particle” etc. solely and/or jointly accumulate the meanings of the biological, biomedical and biochemical agents, cells, molecules, any and all explosive and hazardous agents, aerosol, airborne particles, micro- and nano-particles, liquid/fluid contaminations/particles, gas particles, analytes, etc. Therefore, the use at least one of the terms does not exclude the other meanings for the used term, if otherwise not specified). The same if relevant for the terms, for example, such as “light” and/or “laser” and/or “electromagnetic energy”, etc. [for instance, related to: “source(s)” and/or “beam(s), and/or “radiation(s)”, etc.]. Further, only for simplification of the disclosure the term “biomedical agent” can mostly be presented by the term “agent” or “particle”, and the term “laser”by the term “light”).
Some of them are useful for detection and measurement of the agents illuminated by the light (laser) beam.
There are well known devices and methods of agent (particle) measurement utilizing an open cavity laser and a light scattering techniques. These methods and devices mostly use the optic systems, which are based on the lens use, the same as it described, for example, in U.S. Pat. Nos. 4,140,395; 4,798,465 and 5,495,105.
The other known devices (for example, U.S. Pat. No. 4,606,636) use the mirror systems, e.g., a divergent quadric reflector. Such mirror based devices use a hemi-paraboloidal sphere as a mirror.
In the U.S. Pat. Nos. 4,189,236; 4,523,841; 5,467,189 and 5,515,164) are described the devices (sensors) with non-divergent ellipsoidal mirrors, which also, for example, described in the U.S. Pat. Nos. 5,767,967; 5,946,091; 7,439,855 and 7,573,573, instead of the lens systems or divergent quadric mirrors.
All these devices, mentioned in the prior art above, use the light scattering focalizing methods. Such methods are based on the collection of the scattered light. A light scattering occurs at the first focal point (focus) 8 by particles in the laser beam (in the point of intersection of the particle and light (laser) beam). Considering stochastic dispersion of the scattered light, the devices, mentioned in the above prior art, use mirrors or optics. This is necessary for scattered light collecting and focalizing at the second focal point (focus) 9, where a light detector is placed and intended for scattered light detection [see FIGS. 1, 2, 3, 4 (prior art)].
The FIGS. 1 (prior art)-4 (prior art) (see also FIGS. 1-4 in the U.S. Pat. No. 6,034,769) depict the most common known principles of the light scattering by the open cavity laser: in FIG. 1 (prior art), related to the use of the optics (see the U.S. Pat. Nos. 4,140,395; 4,798,465 and 5,495,105), is shown that the scattered light is collected by the optical system, which is presented by the lenses 10 [also in FIGS. 1-4 (prior art): 1—a device axis, 2—a single light beam axis, 3—a particle flow axis, 5—a plurality of the light detection means (or a large size detection means), 6—the collected scattered light, 8—a first focal point, 9—a second focal points]; in FIG. 2 (prior art) is presented the device, using divergent quadric mirror (U.S. Pat. No. 4,606,636), and from FIG. 2 (prior art) is understandable that the collection of the scattered light is provided by the divergent quadric (paraboloidal) mirror 18; in FIG. 3 (prior art) the counting and measuring devices (sensors), mentioned in the U.S. Pat. Nos. 4,189,236; 4,523,841; 5,467,189 and 5,471,299, use the ellipsoidal mirrors 17; in FIG. 4 (prior art) is presented the particle sensor (measuring and counting device) by U.S. Pat. No. 5,515,164, also using the ellipsoidal mirror for the scattered light collection. This sensor uses specially increased cross-section outlet area of the particle flow. It is understood, that the methods and devices, of the prior art mentioned above, require the use of the scattered light collection means/systems [FIGS. 1-4 (prior arts)]. Such methods and/or devices need to include expensive means and systems. Also, the mentioned above prior art methods and devices have a common deficiency, which is characterized by non-consideration of all scattered light plurality [for example, the unconsidered scattered light 23 in FIGS. 1-4 (prior arts)] and non-precise focalizing of the particle flow [for example, the unfocused scattered light 7 in FIGS. 1-4 (prior arts)].
Some known devices (for example, by U.S. Pat. No. 5,731,875) use a plurality of light emitting low power lasers which provide the elimination of the laser heat-sink, but, it requires to use a plurality of fiber optic stands and the optical element(s) for the focusing of a plurality of light beams, but it will still be a plurality of the beams after collecting optics [not the one (single) enhanced (combined) powerful laser beam] considering that in this patent are used the lasers, which produce the monochromatic coherent light beams (laser beams), which are not combinable into one single powerful beam. Therefore, the particle will intersect not the one single powerful laser beam, but the plurality of the low power laser beams, thereby, creating the incorrect information regarding quantity of the agents (particles) in the assayed (analyzed) specimen.
Thus, the unfocused and/or unconsidered (undetected) scattered light in the mentioned above devices of a prior art creates a light background (light noises) inside such devices, thereby creating incorrectness of the resulting information about the measured agents. Also, such light noises limit the sensitivity of the mentioned devices.
Additionally to the devices using the scattered light collection, there are known the devices using some other optic detection methods, for example, by light splitting, etc.
There are also known some methods and devices useful for detection and measurements of the agents (e.g., airborne agents, etc.) using Raman scattering techniques.
It is well known, that the molecules can be airborne agents, including but not limited to explosives, narcotics, hazardous chemicals, or other chemical or non-chemical species, etc.
As it is well known, the Raman spectroscopy is a technique desired for molecular detection and molecular dynamics studies. Surface Enhanced Raman Scattering (SERS) improves the sensitivity by amplifying the original Raman scattering intensity. The complex plasmon resonance of single nanoparticles and the plasmon correlation with the adjacent nanoparticles are the focus of the biomedical field. The most SERS grounds of nanoparticle combinations related and depend on the size of the particles. It is known, that the SERS spectroscopy is useful to provide a chemical-bond information and biomolecular flat imaging.
The characteristics of the nanoparticles are extremely useful for in vitro and/or in vivo diagnostic analysis and/or imaging. One such characteristic of nanoparticles is the size of the particles.
For example the U.S. Pat. Nos. 7,283,215 and 7,087,444, and the U.S. Patent Publications Nos. 2007/0048746 and 2008/0270042 describe the devices of detection of the airborne agents in microfluidics. It is known, that microfluidics is a field of work that deals with the fluid-based transport of mass, and that the microfluidic channels are generally enclosed and not in direct relation with the surrounding environment (e.g., atmosphere, etc.).
Another U.S. Pat. No. 8,017,408 also describes the method and device of detection of the airborne agents using SERS principles for particle detection and microfluidics, utilizing free-surface fluidics. In general, the disclosure provides a microfluidic platform for real time sensing of volatile airborne agents, such as explosives. The device provides the multiple length scales, ranging from tens of micrometers to a few nanometers. Free-Surface Fluidics (FSF), are used such that one or more fluid surfaces of a fluid flow channel flow are exposed to the surrounding atmosphere, with confinement being caused by surface tension forces operating, typically, on open-channel flows of order depth. This free-surface fluidic (FSF) architecture is incorporated with Surface Enhanced Raman Spectroscopy (SERS) for detection of airborne agents.
The FSF architecture provides at least on of a plurality of surfaces of the microfluidic flow to be exposed to the atmosphere. Since the length scale is on the order of a few microns, a very large surface area is exposed to the atmosphere. This provides automatic injection of airborne molecules into the microfluidic channel.
The disclosure provides a free-surface detection device comprising: a substrate; a fluid flow channel having a first end and a second end located in or on the substrate; at least one free-surface interface region located between the first end and the second end, wherein the free-surface interface region is open on at least one side to atmospheric air comprising an analyte; an excitation area, wherein electromagnetic energy excites a SERS probe in a fluid flowing in the fluid flow channel containing analytes; and a detection area, wherein a scattered light is detected by a detection device, wherein the free-surface interface region is in fluid communication with the excitation and detection areas.
Such methods and devices use the direct detection principles avoiding the delivering and/or collecting optics or mirrors. Also such methods and devices provide the Raman spectroscopy, but do not analyze the quantity and/or concentration of the agents (particles) which are presented at least by two dimensions or preferably by three dimensions.
The disclosure also describes a method for analyte detection comprising: providing a flow of a fluid through a fluid channel in a fluidic device, the fluidic device comprising: a substrate; a fluid flow channel having a first end and a second end located in or on the substrate; at least one free-surface interface region located between the first end and the second end, wherein the free-surface interface region is open on at least one side to atmospheric air comprising an analyte; an excitation area, wherein electromagnetic energy excites a SERS probe in a fluid flowing in the fluid flow channel containing analytes; and a detection area, wherein the scattered light is detected by a detection device, wherein the free-surface interface region is in fluid communication with the excitation and detection areas and wherein analytes in the sample are absorbed into the fluid; a laser source that emits light at the excitation area; and a detector that detects the scattered light from excited SERS probes, contacting the fluid with a SERS probe; and measuring emissions of SERS probes aggregated within the fluid with an analyte, wherein the scattered light is indicative of the presence of analyte in the sample.
More specifically the device comprises a substrate upon which or within which a reservoir or inlet is fluidly connected to an outlet by a fluid channel. The reservoir or inlet is about 40 μm deep (where “μm” means micron). Fluid channel comprises contiguous different regions having a proximal fluid flow region, free-surface interface region and distal fluid flow region. Also the device includes the first region and the detection region, which, when present, are approximate locations in the distal fluid flow region, or in the free-surface interface region. The first regions and detection coincide. During the use of the Raman principles, fluorescent or infrared radiation spectroscopy, regions coincide based upon excitation and emission timing. This prior art invention provides fluidic devices that can drive and guide liquid flow in microfluidic channels to transport biomolecules and living cells.
Such devices, providing the particle direct detection, do not differentiate (distinguish) and/or recognize the particles by their spatial form (shape), and do not provide the counting of particles (e.g., microparticles or nanoparticles, etc.) and/or specific analytes (for instance, in biomolecules and living cells, etc.) in vitro and/or vivo diagnostic testing with respect to the particle spatial size.
There is known the other inventions intended for analysis of a constituent of body fluid using the fluidic channel (e.g., U.S. Pat. No. 8,050,729).
Another known method, also using the scattered light direct detection principles, is described in U.S. Pat. No. 5,085,500. The scattered light in this device is detected by a plurality of light detectors 5 directly with no scattered light optic and/or mirrors collection. In FIG. 5 (prior art) [see also FIG. 5 (prior art) in the U.S. Pat. No. 6,034,769 and FIG. 4a in U.S. Pat. No. 5,085,500] is shown a simplified drawing of the device (U.S. Pat. No. 5,085,500), using the scattered light direct detection method.
It is understood, that the mentioned method and device, require the use of the very large spatial surface (e.g., cylindrical, etc.) of the light detector or sufficient quantity of the light detectors surrounding the focal point. Such method and/or device need to include expensive detection means and systems. Also, the mentioned above method and device have an additional common deficiency, which is characterized by non-consideration of all scattered light [for example, a scattered light 23 in FIG. 5 (prior art)].
The U.S. Pat. No. 6,034,769 describes the method and device for particle direct detection, which is free of the deficiencies of the U.S. Pat. No. 5,085,500 using the scattered light collection principles. The methods and device by U.S. Pat. No. 6,034,769 provide the precise measuring and counting of the micro- and nano-particles.
According to the U.S. Pat. No. 6,034,769, a single light or laser beam of the single light source intersects a particle flow (along axis 3 of the particle flow means 26) within the particle monitoring region in the area of the single light detection means which is placed on a single light beam axis 2. The active area (e.g., photoelement, etc. [not shown]) of the single light detection means 4 (e.g., photodetector/pho-todiode, etc.) of this prior art is fully exposed to the single light beam, as it is shown in FIG. 6 (prior art) [see also FIGS. 6, 7 of the U.S. Pat. No. 6,034,769].
The mentioned hereinabove methods and devices provide one dimension (size) measuring (not spatial, e.g., such as an imaging) of the agents/particles, therefore, such methods and devices can not differentiate/distinguish and/or recognize the form/shape of the analyzed biomedical agents and more particularly can not differentiate/distinguish and/or recognize the form/shape of the analyzed microorganism, cell, molecule and/or analyte in vitro and/or in vivo diagnostic analysis and/or imaging, etc.
Therefore, the mentioned known methods and devices have the described above deficiencies which are eliminated in the improved methods and apparatus for biomedical agent multi-dimension measuring and analysis.
Considering that some microorganisms or cells can have a specific spatial form/shape, for example, an elongated (e.g., “worm”-alike) form or regular or irregular spherical-alike form, etc., the more convenient to recognize and identify (verify) the biological organisms by their forms, for example, before the complex analysis of their spectroscopic, chemical, magnetic, etc. characteristics. Also, it is more precise and authentic to recognize, select and sort the particles by their, for example, three-dimensions (X, Y, Z), but not only by the particle's single size (e.g., X-axis measurement). Additionally, the improved method and apparatus provide possibility to spatially depict/display the multi-dimension measured particles.
While the mentioned above prior art fulfill their respective, particular objectives and requirements, the mentioned prior art inventions do not disclose, teach and/or suggest the methods and apparatus for biomedical agent multi-dimension measuring and analysis including the steps (and their sequence) of the methods and elements (components/parts) of the apparatus providing the possibility of the differentiation/distinguishing and/or recognition of the form/shape of the analyzing microorganism, cell, molecule and/or analyte in vitro and/or in vivo diagnostic analysis and/or imaging, etc.
Those skilled in the art will readily observe that numerous modifications and advantages of the improved methods and apparatus for biomedical agent multi-dimension measuring and analysis may be made while retaining the teachings of the invention.
Thus, the known prior art do not provide the efficient, precise, and convenient methods and apparatus for biomedical agent multi-dimension measuring and analysis according to the present invention substantially departs from the devices of the prior art.